玻璃化冷冻对小鼠窦前卵泡体外培养及卵子骨架的影响  

EFFECTS OF VITRIFICATION ON EARLY PREANTRAL MOUSE FOLLICLES IN VITRO CULTURE AND CYTOSKELETON IN MICE

作　　者：邹长林[1] 葛红山[2] 陈雅[3] 张巍[3] 薛亚梅[2] 

机构地区：[1]温州医学院附属第一医院流行病学教研室,浙江温州325000 [2]温州医学院附属第二医院生殖健康中心,浙江温州325000 [3]温州医学院附属第一医院生殖医学中心,浙江温州325000

出　　处：《河北医科大学学报》2008年第5期681-684,805,共5页Journal of Hebei Medical University

基　　金：浙江省自然基金资助(编号Y204272)

摘　　要：目的分析玻璃化冷冻对小鼠窦前卵泡体外生长、发育及成熟后卵子细胞骨架的影响。方法超薄型开口拉细麦管（super open pulled straw,SOPS）玻璃化冷冻12～14日龄小鼠窦前卵泡,解冻复苏后,将卵泡移入α基础培养液（αmini mumessential medium,α-MEM）＋10%胎牛血清（fetal calf serum,FCS）＋0.075I U/mL卵泡刺激素＋胰岛素-转铁蛋白-硒＋10mIU/mL人绒毛膜促性腺激素（human chorionic gonadotrophin,HCG）中培养,隔日换一半培养液。第12d将卵泡转入α-MEM＋10%FCS＋1I U/mL HCG＋5ng/mL表皮细胞生长因子,16～18h后收集卵子。免疫荧光技术测定卵子纺锤体和染色体结构。结果卵泡解冻后成活率为92.0%,卵泡体外生长速率、卵泡腔形成与新鲜卵泡相比均没有明显改变（P〉0.05）。冷冻对卵泡成熟后卵子骨架和染色体没有显著影响（P〉0.05）。结论SOPS玻璃化冷冻技术是一种高效可靠的卵泡冷冻方法。Objective To determine potential cryopreserved damages on the capacity of in vitro growth of early preantral mouse follicles. Methods Early preantral mouse follicles were vitrified using super open pulled straw（SOPS）. After thawed, follicles were cultured in medium for up to 13 days, change half medium every other day, recorded follicle sizes and antral cavity formation. Matured oocytes collected after culture were fixed for immunofluorescence. Results The survival rate of vitrified-thawed follicles was 92.0%： There were no significant differences on the growth of follicle in vitro and the rate of antral cavity formation between vitrified-thawed and fresh follicles. The rates of both spindle and chromosomes with normal configuration in vitrified-thawed and fresh groups were no different. Conclusion The method of vitrified preantral follicles Using SOPS is efficient and credible.

关 键 词：卵泡 冷冻 小鼠 

分 类 号：R321-33[医药卫生—人体解剖和组织胚胎学]